ID 112045
Title Alternative
Cell Biological Approaches
Author
Keywords
trichloroacetic acid(TCA)fixation
Rho
cytokinesis
epithelial sheets
Content Type
Journal Article
Description
Our cell biological approaches are introduced in this article. First, we show development of a new fixation protocol using trichloroacetic acid (TCA) that is ideal for immunofluorescence microscopy detecting cellular distribution of phosphorylated ERM (ezrin/radixin/moesin) proteins. TCA denatures proteins differently from organic solvents or aldehydes and can be a good alternative to conventional fixatives when immunostaining is not successful. Second, using this new fixation protocol, the small GTPase, Rho was found to be successfully immunostained and this led to understanding of molecular mechanism for determination of the position of the cleavage furrow at cytokinesis through microtubules of the mitotic apparatus. Rho activator is translocated to the presumptive furrow region through microtubules, then, Rho is accumulated and activated there leading to myosin II contractility for furrowing. Third, we show the cell ablation using laser beam during microscopic observation. This enables us to analyze cell responses to the death of neighboring cells in epithelial sheets at early stages.
Journal Title
Shikoku Acta Medica
ISSN
00373699
NCID
AN00102041
Publisher
徳島医学会
Volume
73
Issue
5-6
Start Page
241
End Page
248
Sort Key
241
Published Date
2017-12-25
FullText File
language
jpn
TextVersion
Publisher
departments
Medical Sciences